KR102237888B1 - Bivalent inhibitors of iap proteins and therapeutic methods using the same - Google Patents

Abstract

Inhibitors of the IAP protein and compositions containing the same are disclosed. Also disclosed is a method of using an IAP protein inhibitor in the treatment of diseases and conditions, such as cancer, in which inhibition of the IAP protein benefits.

Description

Bivalent inhibitor of IAP protein and treatment method using the same {BIVALENT INHIBITORS OF IAP PROTEINS AND THERAPEUTIC METHODS USING THE SAME}

Government support

The present invention was made with U.S. government support under approval numbers CA127551 and CA109025 awarded by the National Institutes of Health. The US government has certain rights in this invention.

Field of invention

The present invention relates to divalent inhibitors of inhibitors of apoptosis proteins (IAPs), and methods of treatment for treating conditions and diseases in which inhibition of IAP proteins provides a benefit. The inhibitors of the present invention bind to IAP proteins including cIAP1, cIAP2 and XIAP with very high affinity to induce apoptosis in human cancer cell lines, thereby enhancing the antitumor activity of other anticancer drugs.

Apoptosis or programmed cell death is an important cellular process for homeostasis, normal development, host defense, and inhibition of oncogenesis. Incomplete regulation of apoptosis is associated with cancer ^{(1), (3)} and many human diseases ^(1), and resistance to apoptosis is currently recognized as a characteristic of cancer. ⁽⁴⁾ As a result, targeting of key apoptosis modulators is emerging as an attractive strategy for the development of new approaches for the treatment of human cancer. ^(One)

Most current cancer therapies, including chemotherapeutic agents, radiation, and immunotherapy, indirectly induce apoptosis in cancer cells. Thus, the failure of cancer cells to perform an apoptosis program due to defects in the normal apoptotic machinery is often associated with an increase in resistance to chemotherapy, radiation or immunotherapy-induced apoptosis. This congenital or acquired resistance of human cancer to current therapies due to apoptosis defects is a major problem in current cancer therapy.

To improve the survival and quality of life of cancer patients, current and future efforts in the design and development of new molecular target-specific anticancer therapies include strategies to specifically target cancer cells that are resistant to apoptosis. . In this regard, targeting negative regulators that play a pivotal role in directly inhibiting apoptosis in cancer cells represents a very promising therapeutic strategy for designing new anticancer drugs.

One class of key negative regulators of apoptosis are inhibitors of apoptosis proteins (IAPs). This class includes proteins such as XIAP, cIAP1, cIAP2, ML-IAP, HIAP, KIAP, TSIAP, NAIP, Survivin, Livin, ILP-2, Apollon and BRUCE. The IAP protein strongly inhibits cancer cell apoptosis induced by many different apoptosis stimuli, including chemotherapeutic agents, radiation and immunotherapy.

Although its role is not limited to the regulation of apoptosis, ^(7),(8) IAP proteins are a class of major apoptosis regulators and are characterized by the presence of one or more BIR (baculovirus IAP repeat) domains. ^{Among the (5)-(6)} IAPs, cells IAP1 (cIAP1) and cIAP2 play a major role in the regulation of death-receptor mediated apoptosis, while X-linked IAP (XIAP) is the three cysteines that are important for the performance of apoptosis. It inhibits both death-receptor mediated and mitochondrial mediated apoptosis by binding to and inhibiting the proteases caspase-3/7 and caspase-9. ⁽⁵⁾ These IAP proteins are highly overexpressed in both cancer cell lines and human tumor tissues and low in normal cells and tissues. ⁽⁹⁾ Intensive studies have demonstrated that overexpression of the IAP protein makes cancer cells resistant to induction of apoptosis by various anticancer drugs. ^(10)-(12) A detailed discussion of the IAP protein and its role in cancer and apoptosis is described in US Pat. No. 7,960,372, which is incorporated herein by reference. Thus, targeting one or more of these IAP proteins is a promising therapeutic strategy for the treatment of human cancer. ^(10)-(12)

Studies have revealed that peptide-based inhibitors are useful tools for elucidating the anti-apoptotic function of IAP and the role of IAP in the response of cancer cells to chemotherapeutic agents. However, peptide-based inhibitors have inherent limitations as useful therapeutic agents, including poor cell permeability and poor in vivo stability. In published studies using Smac-based peptide inhibitors, the peptide had to be fused to a carrier peptide in order for it to be relatively cell-permeable.

Small molecule inhibitors of IAP proteins are also known. For example, U.S. Patent Publication No. 2005/0197403 and U.S. Patent No. 7,960,372 disclose dimeric Smac mimetic compounds, each of which is incorporated herein by reference in its entirety.

Despite the discovery of small molecule inhibitors of IAP proteins, the design of potent non-peptide inhibitors of IAP proteins remains a significant challenge in modern drug discovery. Accordingly, there is still a need in the art for IAP inhibitors with physical and pharmacological properties that allow the use of the inhibitor in therapeutic applications. The present invention provides compounds designed to bind to IAP protein and inhibit IAP protein activity.

It is generally accepted that the failure of cancer cells or their supporting cells to undergo apoptosis in response to genetic damage or exposure to inducers of apoptosis (such as chemotherapeutic agents and radiation) is a major factor in the onset and progression of cancer. Induction of apoptosis in cancer cells or their supporting cells (eg, neovascular cells in the tumor vascular system) is today considered a universal mechanism of action for virtually all effective cancer treatment drugs and radiation therapy. One reason cells do not undergo apoptosis is the increased expression and accumulation of IAP.

Accordingly, the present invention relates to inhibitors of IAP proteins, compositions comprising the inhibitors, and methods of using the inhibitors in the therapeutic treatment of conditions and diseases in which inhibition of IAP protein activity provides a benefit. The compounds of the present invention are potent inhibitors of IAP protein activation and induce apoptosis in cancer cells.

^{1 is an average tumor volume (mm 3} ) vs. anti-tumor activity of Examples 2 and 24 in the MDA-MB-231 xenograft model in nude mice. It is a plot of days after transplantation.

More particularly, the present invention relates to compounds having the structural formula (I), or to a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof.

<Formula I>

,

,

및 -SO₂-로 이루어진 군으로부터 선택되고;In the above formula, X is

,

,

And -SO ₂ -;

Y is selected from the group consisting of -NH-, -O-, -S- and absent;

,

(여기서 고리 A는 C_4-8 지방족 고리임),

및

로 이루어진 군으로부터 선택되며, 여기서 B 고리는 아릴 또는 질소 원자-함유 헤테로아릴이고 B 고리는 임의로 치환되고;R is

,

(Where ring A is a C _4-8 aliphatic ring),

And

Is selected from the group consisting of, wherein ring B is an aryl or nitrogen atom-containing heteroaryl and ring B is optionally substituted;

,

,

,

,

(여기서 Z는 O, S 또는 NH임) 및

(여기서 n은 0, 1 또는 2임)로 이루어진 군으로부터 선택되며, 여기서 B 고리는 아릴 또는 질소 원자-함유 헤테로아릴이고

고리는 임의로 치환된다.R ₁ is -(CH ₂ ) _4-10 -,

,

,

,

,

(Where Z is O, S or NH) and

(Where n is 0, 1 or 2) is selected from the group consisting of, wherein the B ring is an aryl or nitrogen atom-containing heteroaryl

The ring is optionally substituted.

In one embodiment, the invention provides compounds that inhibit the activity of an IAP protein and increase the susceptibility of a cell to apoptosis inducing agents such as chemotherapeutic agents and radiation therapy.

In other embodiments, the compounds of the invention are used in a method of inducing apoptosis in a cell and sensitizing a cell to an apoptosis inducing agent.

In another embodiment, the invention provides a method of treating a condition or disease by administering a therapeutically effective amount of a compound of structural formula (I) to an individual in need thereof. The disease or condition of interest, such as cancer, is treatable by inhibition of the IAP protein. Accordingly, the compounds of the present invention are useful for the treatment and amelioration of disorders responsive to the induction of apoptotic cell death, for example, hyperproliferative diseases, such as those characterized by dysregulation of apoptosis, including cancer. In certain embodiments, the compounds can be used to treat and ameliorate cancer characterized by resistance to cancer therapy (eg, chemo resistance, radiation resistance, hormone resistance, etc.). In other embodiments, the compounds of the present invention can be used to treat a hyperproliferative disease characterized by overexpression of IAP.

Another embodiment of the invention is a composition comprising (a) an IAP inhibitor of structural formula (I) and (b) an excipient and/or a pharmaceutically acceptable carrier useful for treating a disease or condition in which inhibition of the IAP protein provides a benefit. Is to provide.

Another embodiment of the invention is the use of a composition comprising a compound of structural formula (I) and a second therapeutically active agent in a method of treating a disease or condition for which inhibition of an IAP protein in an individual would benefit.

In a further embodiment, the invention provides the use of a composition comprising an IAP protein inhibitor of structural formula (I) and an optional second therapeutic agent for the manufacture of a medicament for the treatment of a disease or condition of interest, such as cancer.

Another embodiment of the invention comprises (a) a container, (b1) a packaged composition comprising an IAP protein inhibitor of structural formula I, and optionally, (b2) a second therapeutic agent useful for the treatment of a disease or condition of interest. To provide a kit for human pharmaceutical use comprising a packaged composition and (c) a package insert containing instructions for the use of the composition, or compositions administered simultaneously or sequentially, in the treatment of a disease or condition.

The IAP protein inhibitor of structural formula I and the second therapeutic agent can be administered together as a single-unit dose or separately as a multi-unit dose, wherein the IAP inhibitor of structural formula I is administered before the second therapeutic agent or vice versa. . It is envisioned that one or more doses of an IAP inhibitor of structural formula I and/or one or more doses of a second therapeutic agent may be administered.

In one embodiment, the IAP protein inhibitor of structural formula (I) and the second therapeutic agent are administered simultaneously. In a related embodiment, the IAP protein inhibitor of structural formula I and the second therapeutic agent are administered from a single composition or from separate compositions. In a further embodiment, the IAP protein inhibitor of structural formula (I) and the second therapeutic agent are administered sequentially. The IAP protein inhibitors of structural formula I as used in the present invention may be administered in an amount of about 0.005 to about 500 milligrams per dose, about 0.05 to about 250 milligrams per dose, or about 0.5 to about 100 milligrams per dose.

Detailed description of preferred embodiments

The invention is described in connection with a preferred embodiment. However, it will be apparent that the present invention is not limited to the disclosed embodiments. Given the description of embodiments of the present invention herein, it is understood that various modifications may be made by those skilled in the art. Such modifications are covered by the following claims.

Smac/DIABLO (second mitochondrial-derived activator of caspase or direct IAP binding protein, with low pi) is a protein released from mitochondria in response to apoptosis stimulation, and functions as endogenous inhibitors of cIAP1, cIAP2 and XIAP. ^{(14), (15)} The interaction between Smac and IAP is mediated by the N-terminal AVPI tetrapeptide motif in Smac and one or more BIR domains in these IAP proteins. ^{(16), (17)} Smac is a homodimer that binds to both the BIR2 and BIR3 domains in XIAP and antagonizes the inhibition of XIAP against caspase-3/-7 and caspase-9. ⁽¹⁸⁾ In comparison, Smac binds only to the BIR3 domains in cIAP1 and cIAP2 and ⁽¹⁹⁾ induces rapid proteolysis in cells. ⁽²⁰⁾ Through two distinct mechanisms, Smac is a highly efficient antagonist of these three IAP proteins.

The crystal and NMR structure of XIAP BIR3 complexed with Smac protein or Smac peptide shows that the AVPI tetrapeptide motif in Smac binds to the well-defined surface groove in XIAP, and this interaction is for the design of small molecule XIAP inhibitors. Present attractive areas. Several classes of small molecule Smac mimetics, by the use of AVPI tetrapeptide as the ^{(16)-(18) lead structure, have been designed as antagonists of XIAP and cIAP1/2. (21)-(38)} Two different types of Smac mimetics have been designed. ^(21)-(23) The first type is designed to mimic a single AVPI binding motif and is called a monovalent Smac mimic. ^{The second type (21)-(23)} is a divalent Smac mimetic, consisting of two AVPI mimetics tethered through a linker to mimic the dimeric form of the Smac protein. ^(21)-(23)

One advantage of the monovalent Smac mimetic as a potential drug is oral bioavailability, but its drawback is its modest potency in antagonizing full-length XIAP in functional assays. The main advantage of the bivalent Smac mimetic is that it is a much more potent antagonist of XIAP than the monovalent Smac mimic by jointly targeting both the BIR2 and BIR3 domains within XIAP. ⁽³⁰⁾ Bivalent Smac mimetics are typically 2-3 times more potent than their monovalent Smac mimetic counterparts in inducing apoptosis in cancer cells. ⁽²¹⁾ Currently, three monovalent and two divalent Smac mimetics are in clinical trials for the treatment of human cancer. ⁽²¹⁾

Since the divalent Smac mimetic is significantly more potent than the monovalent Smac mimetic in targeting XIAP and cIAP1/2 in inducing apoptosis of cancer cells in vitro and in vivo and in inhibiting tumor growth, the present invention Divalent compounds are designed for use in the treatment of cancer and other diseases and conditions mediated by IAP protein activity.

The term “IAP protein” as used herein includes, but is limited to, XIAP, cIAP-1, cIAP-2, ML-IAP, HIAP, TSIAP, KIAP, NAIP, Survivin, Livin, ILP-2, Apollon and BRUCE. Refers to any known member of the inhibitors of the apoptotic protein family.

The term “overexpression of IAP” as used herein expresses an elevated level (eg, an abnormal level) of an mRNA encoding an IAP protein(s) and/or a basal level of an mRNA encoding an IAP protein or an IAP protein. Refers to an elevated level of IAP protein(s) in a cell compared to a similar corresponding non-pathological cell with a basal level of. Methods for detecting the level of the mRNA encoding the IAP protein or the level of the IAP protein in a cell include, but are not limited to, Western blotting using an IAP protein antibody, an immunohistochemical method, and a nucleic acid amplification method or direct RNA detection. Does not. As important as the absolute level of the IAP protein in the cell is to determine if it overexpresses the IAP protein, and also for other apoptosis-promoting signaling molecules (e.g., apoptosis-promoting Bcl-2 family proteins) within these cells. It is the relative level of protein. If the balance of these two would be sufficient for the apoptosis-promoting signaling molecule to carry out the apoptosis program and kill the cell without the level of the IAP protein, the cell would be dependent on the IAP protein for its survival. In such cells, exposure to an inhibitory effective amount of an IAP protein inhibitor will be sufficient to cause the cell to undergo an apoptotic program and die. Thus, the term “overexpression of an IAP protein” also refers to a cell that undergoes apoptosis in response to an inhibitory effective amount of a compound that inhibits the function of the IAP protein, due to the relative levels of the apoptosis-promoting signal and the anti-apoptotic signal.

The term "disease or condition in which the inhibition of an IAP protein provides a benefit" refers to a condition in which the action of an IAP protein and/or an IAP protein is important or necessary, for example for the onset, progression or expression of such a disease or condition, or an IAP protein inhibitor. It relates to a disease or condition known to be treated by. Examples of such conditions include, but are not limited to, cancer. One of skill in the art can readily determine for any particular cell type whether a compound treats a disease or condition mediated by an IAP protein, e.g., by assays that can be conveniently used to assess the activity of a particular compound. have.

The term “second therapeutic agent” refers to a therapeutic agent that is different from the IAP inhibitor of structural formula (I) and is known to treat the disease or condition of interest. For example, if the cancer is a disease or condition of interest, the second therapeutic agent may be a known chemotherapy drug such as Taxol or radiation.

The term “disease” or “condition” generally refers to a disorder and/or condition that is considered to be a pathological condition or function and can manifest itself in the form of specific signs, symptoms and/or dysfunction. As demonstrated below, the compounds of structural formula I are potent inhibitors of the IAP protein and can be used to treat diseases and conditions where inhibition of the IAP protein provides benefits.

As used herein, the terms “treat,” “treating,” “treatment,” and the like refer to eliminating, reducing or ameliorating a disease or condition and/or symptoms associated therewith. Although not excluded, treating a disease or condition does not require complete elimination of the disease, condition, or symptoms associated therewith. As used herein, the terms "treat," "treating," "treatment," and the like refer to the recurrence of a disease or condition or a recurrence of a disease or condition in a subject that is at risk or susceptible to, or And “prophylactic treatment”, which refers to reducing the probability of recurrence of a pre-controlled disease or condition. The terms “treat” and synonyms contemplate administering a therapeutically effective amount of a compound of the invention to an individual in need of such treatment.

Within the meaning of the present invention, “treatment” includes prevention of recurrence or prevention of modalities, as well as treatment of acute or chronic signs, symptoms and/or dysfunction. Treatment can be set symptomatic, for example to suppress symptoms. This can be done over a short period of time, can be set over an intermediate period, or can be a long-term treatment, for example in the context of maintenance therapy.

The terms “sensitize” and “sensitize” as used herein refer to the administration of a first agent (eg, a compound of structural formula I), so that the animal or cells in the animal For example, it refers to making it more susceptible or more responsive to promoting or delaying aspects of cellular function, including, but not limited to, cell division, cell growth, proliferation, invasion, angiogenesis or apoptosis. The sensitizing effect of the first agent on the target cells is the intended biological effect observed upon administration of the second agent with and without administration of the first agent (e.g., cell growth, proliferation, invasion, angiogenesis or apoptosis. Including, but not limited to, promoting or delaying aspects of cellular function). The response of the sensitized cell is at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% than the response in the absence of the first agent. , At least 100%, at least 150%, at least 200%, at least 350%, at least 300%, at least 350%, at least 400%, at least 450%, or at least 500%.

As used herein, the term “hyperproliferative disease” refers to any condition in which a localized population of proliferating cells in an animal is not controlled by conventional limitations of normal growth. Examples of hyperproliferative disorders include, but are not limited to, tumors, neoplasms, lymphomas, and the like. A neoplasm is called benign if it does not undergo invasion or metastasis, and malignant if any of these are true. "Metastatic" cells mean that the cells are able to invade and destroy surrounding body structures. Hyperplasia is a form of cell proliferation that involves an increase in the number of cells in a tissue or organ without significant alteration in structure or function. Metabolism is a form of controlled cell growth in which one type of fully differentiated cell replaces another type of differentiated cell.

Pathological growth of activated lymphoid cells often leads to autoimmune disorders or chronic inflammatory conditions. As used herein, the term “autoimmune disorder” refers to any condition in which an organism produces antibodies or immune cells that recognize molecules, cells or tissues of the organism itself. Non-limiting examples of autoimmune disorders include autoimmune hemolytic anemia, autoimmune hepatitis, Berger's disease or IgA nephropathy, celiac sprue, chronic fatigue syndrome, Crohn's disease, dermatitis, fibromyalgia, graft versus host disease, Graves disease, Hashimoto's thyroiditis, idiopathic thrombocytopenia purpura, lichen planus, multiple sclerosis, myasthenia gravis, psoriasis, rheumatic fever, rheumatoid arthritis, scleroderma, Sjogren's syndrome, systemic lupus erythematosus, type 1 diabetes, ulcerative colitis, vitiligo and others. Includes.

The term “neoplastic disease” as used herein refers to any abnormal growth of cells that are benign (non-cancerous) or malignant (cancerous).

The term “anti-neoplastic agent” as used herein refers to any compound that delays the proliferation, growth or spread of a targeted (eg, malignant) neoplasm.

The term “apoptosis-modulator” as used herein refers to an agent that is involved in the regulation of apoptosis (eg, inhibition, decrease, increase, promote). Examples of apoptosis-modulating agents include proteins comprising death domains such as, but not limited to, Fas/CD95, TRAMP, TNF RI, DR1, DR2, DR3, DR4, DR5, DR6, FADD and RIP. Other examples of apoptosis-modulating agents include antibodies to TNFα, Fas ligand, Fas/CD95 and other TNF family receptors, TRAIL (also known as Apo2 ligand or Apo2L/TRAIL), agonists of TRAIL-R1 or TRAIL-R2 (e.g. For example, monoclonal or polyclonal agonistic antibodies), Bcl-2, p53, BAX, BAD, Akt, CAD, PI3 kinase, PP1 and caspase proteins, but are not limited thereto. Modulators broadly include agonists and antagonists of TNF family receptors and TNF family ligands. Apoptosis-modulating agents may be soluble or may be membrane bound (eg ligand or receptor). Preferred apoptosis-modulating agents are apoptosis inducing agents, such as TNF or TNF-related ligands, in particular TRAMP ligand, Fas/CD95 ligand, TNFR-1 ligand or TRAIL.

The term “dysregulation of apoptosis” as used herein refers to any abnormality in the ability (eg, tendency) of a cell to undergo cell death through apoptosis. Dysregulation of apoptosis can be, for example, autoimmune disorders (e.g., systemic lupus erythematosus, rheumatoid arthritis, graft-versus-host disease, myasthenia gravis or Sjogren syndrome), chronic inflammatory conditions (e.g., psoriasis, asthma or Crohn's disease), hyperproliferative disorders (e.g., tumors, B cell lymphomas or T cell lymphomas), viral infections (e.g., herpes, papillomas or HIV) and other conditions such as osteoarthritis and atherosclerosis. Associated with or caused by. It should be noted that if the dysregulation is caused by or is associated with a viral infection, the viral infection may or may not be detected at the time dysregulation occurs or it is observed. That is, virus-induced dysregulation can occur even after the disappearance of symptoms of viral infection.

The term “therapeutically effective amount” or “effective dose” as used herein refers to an individual in need of treatment of the condition or disease with the active ingredient(s) for the treatment of the condition or disease of interest when administered by the method of the present invention. It refers to the amount of active ingredient(s) sufficient to effectively deliver to. In the case of cancer or other proliferative disorders, a therapeutically effective amount of an agent can reduce (ie, to some extent delay and preferably stop) undesired cell proliferation and/or; Reduce the number of cancer cells and/or; Can reduce tumor size and/or; Inhibit (ie to some extent and preferably arrest) cancer cell invasion into peripheral organs; Inhibit (ie to some extent and preferably arrest) tumor metastasis; Inhibit tumor growth to some extent and/or; Reduce IAP protein signaling in target cells and/or; May increase the duration of survival and/or; One or more of the symptoms associated with cancer to some extent, at least 5%, preferably at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, At least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%. The administered compound or composition may be cytostatic and/or cytotoxic to the extent that it is capable of preventing the growth of existing cancer cells and/or killing existing cancer cells.

The term “container” means any storage and enclosure suitable for storing, transporting, dispensing and/or handling pharmaceutical products.

The term “insert” means information accompanying a pharmaceutical product that provides a description of how the product is administered, along with the safety and efficacy data necessary to enable physicians, pharmacists, and patients to make informed decisions regarding the use of the product. do. Package inserts are generally regarded as "markers" for pharmaceutical products.

“Co-administration,” “administered in combination,” “concurrent administration” and similar phrases mean that two or more agents are administered concurrently to a subject to be treated. “Concurrently” means that each agent is administered sequentially in any order at the same time or at different time points in time. However, when not administered simultaneously, this means that they are administered to the subject in close enough time in turn to provide the desired therapeutic effect and act in concert. For example, the IAP protein inhibitor of structural formula (I) can be administered sequentially in any order simultaneously with the second therapeutic agent or at different time points in time. The IAP protein inhibitor of the present invention and the second therapeutic agent can be administered separately in any suitable form and by any suitable route. It is understood that when the IAP protein inhibitor of the present invention and the second therapeutic agent are not administered concurrently, it can be administered to a subject in need thereof in any order. For example, the IAP protein inhibitor of the present invention is administered prior to administration of the second therapeutic modality (e.g., radiotherapy) (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks or 12 weeks ago), in parallel , Or thereafter (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week , After 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks or 12 weeks) it can be administered to an individual in need thereof. In various embodiments, the IAP protein inhibitor of structural formula (I) and the second therapeutic agent are 1 minute apart, 10 minute intervals, 30 minute intervals, less than 1 hour intervals, 1 hour intervals, 1 hour to 2 hours intervals, 2 hours to 3 hours. Intervals, 3 to 4 hours intervals, 4 to 5 hours intervals, 5 to 6 hours intervals, 6 to 7 hours intervals, 7 to 8 hours intervals, 8 to 9 hours intervals, 9 to 10 hours intervals , 10 to 11 hours, 11 to 12 hours, 24 hours or less, or 48 hours or less. In one embodiment, the components of the combination therapy are administered at intervals of 1 minute to 24 hours.

The use of singular terms and similar designations in the context in which the invention is described (especially in the context of the claims) is to be construed as encompassing both the singular and the plural unless otherwise indicated. Reference to a range of values herein is intended to be provided simply as an abbreviated method of referring individually to each distinct value falling within the range, unless otherwise indicated herein, each distinct value being used herein as it is individually. Is incorporated herein by reference. The use of any and all examples, or illustrative expressions (eg, “such as”) provided herein is intended to better illustrate the invention, and unless otherwise stated in the claims, without regard to the scope of the invention. It is not a limitation. No expression in the specification is to be construed as representing any unclaimed element as essential to the practice of the present invention.

The present invention relates to compounds of structural formula (I) which are mimetics of Smac and function as inhibitors of the IAP protein. The compounds of the present invention induce apoptosis by sensitizing cells to agents inducing apoptosis, and in some cases by themselves inhibiting the IAP protein. Accordingly, the present invention relates to a method of sensitizing a cell to an agent inducing apoptosis and a method of inducing apoptosis in a cell, comprising contacting the cell with a compound of structural formula (I) alone or with an agent for inducing apoptosis. The invention further relates to a method of treating or ameliorating a disorder in an animal responsive to induction of apoptosis comprising administering to the animal a compound of structural formula I and an agent inducing apoptosis. Such disorders include those characterized by dysregulation of apoptosis and those characterized by overexpression of the IAP protein.

The present invention relates to potent inhibitors of the IAP protein. The IAP protein inhibitor of the present invention binds XIAP, cIAP1 and cIAP2 with low to nanomolar affinity, and is highly effective in antagonizing XIAP in a cell-free functional assay, a non-peptidic, divalent Smac mimetic. to be. The compounds of the present invention effectively induce the degradation of cIAP1 and cIAP2 in cancer cells at low concentrations, activate caspase-3 and -8, and cleave PARP. The compounds of the present invention have a _{low IC 50} for inhibition of cell growth in both the MDA-MB-231 and SK-OV-3 cell lines.

The IAP protein inhibitors of the present invention are thus useful for such treatment in subjects in need of treatment of undesired proliferative cells, including cancer and precancer. Also provided is a method of treating a subject with undesired proliferative cells comprising administering a therapeutically effective amount of a compound of the invention to a subject in need thereof. In addition, proliferation of undesired proliferative cells, such as cancer and precancer, in a subject, comprising administering to a subject at risk of developing a condition characterized by undesired proliferative cells, a therapeutically effective amount of a compound of structural formula (I). Methods of prevention are provided. In some embodiments, the compound of structural formula I decreases the proliferation of unwanted cells by inducing apoptosis in these cells.

The present invention relates to an IAP protein inhibitor having the structural formula (I), or to a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof.

<Formula I>

,

,

및 -SO₂-로 이루어진 군으로부터 선택되고;In the above formula, X is

,

,

And -SO ₂ -;

Y is selected from the group consisting of -NH-, -O-, -S- and absent;

,

(여기서 고리 A는 C_4-8 지방족 고리임),

및

로 이루어진 군으로부터 선택되며, 여기서 B 고리는 아릴 또는 질소 원자-함유 헤테로아릴이고 B 고리는 임의로 치환되고;R is

,

(Where ring A is a C _4-8 aliphatic ring),

And

Is selected from the group consisting of, wherein ring B is an aryl or nitrogen atom-containing heteroaryl and ring B is optionally substituted;

,

,

,

,

(여기서 Z는 O, S 또는 NH임) 및

(여기서 n은 0, 1 또는 2임)로 이루어진 군으로부터 선택되며, 여기서 B 고리는 아릴 또는 질소 원자-함유 헤테로아릴이고

고리는 임의로 치환된다.R ₁ is -(CH ₂ ) _4-10 -,

,

,

,

,

(Where Z is O, S or NH) and

(Where n is 0, 1 or 2) is selected from the group consisting of, wherein the B ring is an aryl or nitrogen atom-containing heteroaryl

The ring is optionally substituted.

As used herein, the term “C _4-8 aliphatic ring” is unsubstituted or refers to 1 to 3 groups such as C _1-4 alkyl, halo, trifluoromethyl, trifluoromethoxy, hydroxy, alkoxy, Refers to cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl substituted with nitro, cyano, alkylamino or amino groups.

The term “alkyl” as used herein refers to straight and branched saturated C _1-10 hydrocarbon groups, non-limiting examples of which include methyl, ethyl, and straight and branched propyl, butyl, pentyl, hexyl, heptyl, octyl, Nonyl and decyl groups. The term C _n means that the alkyl group has “n” carbon atoms.

를 지칭한다. "C_3-6시클로알킬렌"은 비치환되거나, 또는 1 내지 3개의 기, 예를 들어 C_1-4알킬, 할로, 트리플루오로메틸, 트리플루오로메톡시, 히드록시, 알콕시, 니트로, 시아노, 알킬아미노 또는 아미노 기로 치환될 수 있다.The term “C _3-6 cycloalkylene” refers to a disubstituted cycloalkane having 3 to 6 carbon atoms, eg

Refers to. "C _3-6 Cycloalkylene" is unsubstituted or represents 1 to 3 groups such as C _1-4 alkyl, halo, trifluoromethyl, trifluoromethoxy, hydroxy, alkoxy, nitro, cyano It may be substituted with no, alkylamino or amino groups.

The term "alkenyl" is defined identically to "alkyl" except that it contains a carbon-carbon double bond, for example ethenyl, propenyl and butenyl.

As used herein, the term “halo” is defined as fluoro, chloro, bromo and iodo.

The term "hydroxy" is defined as -OH.

The term "alkoxy" is defined as -OR where R is alkyl.

The term "amino" is defined as -NH _2, The term "alkylamino" is more than 1 R alkyl Claim 2 R is defined as alkyl or hydrogen, the -NR _2.

The term "nitro" is defined as _{-NO 2.}

The term "cyano" is defined as -CN.

The term "trifluoromethyl" is defined as _{-CF 3.}

The term "trifluoromethoxy" is defined as _{-OCF 3.}

The term “aryl” as used herein refers to a monocyclic or polycyclic aromatic group, preferably a monocyclic or bicyclic aromatic group, such as phenyl or naphthyl. Unless otherwise indicated, an aryl group is unsubstituted or, for example, halo, alkyl, alkenyl, -OCF ₃ , _{-NO 2} , -CN, -NC, -OH, alkoxy, amino, alkylamino, -CO ₂ H, -CO ₂ alkyl, alkynyl, cycloalkyl, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, silyl, alkylthio, sulfonyl, sulfonamide, aldehyde, heterocycloalkyl, trifluoro It may be substituted with one or more, and in particular, 1 to 4 groups independently selected from romethyl, aryl and heteroaryl.

The term “heteroaryl” as used herein refers to a monocyclic or bicyclic ring system containing one or two aromatic rings and containing one or more and up to four nitrogen atoms in the aromatic ring. Unless otherwise indicated, a heteroaryl group is unsubstituted or, for example, halo, alkyl, alkenyl, -OCF ₃ , _{-NO 2} , -CN, -NC, -OH, alkoxy, amino, alkylamino, -CO ₂ H , -CO ₂ alkyl, alkynyl, cycloalkyl, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, silyl, alkylthio, sulfonyl, sulfonamide, aldehyde, heterocycloalkyl, tri It may be substituted with one or more and in particular 1 to 4 substituents selected from fluoromethyl, aryl and heteroaryl.

를 지칭한다. 용어 "헤테로아릴렌"은 유사하게 정의된다.The term “arylene” refers to a bidentate aryl group that serves to bond and link two different groups, eg

Refers to. The term “heteroarylene” is defined similarly.

Non-limiting examples of aryl groups are as follows.

Non-limiting examples of heteroaryl groups are as follows.

The compounds of structural formula (I) inhibit IAP proteins and are useful for treating a variety of diseases and conditions. In particular, the compounds of structural formula (I) are used in methods of treating diseases or conditions for which inhibition of IAP protein benefits, such as cancer, autoimmune disorders and chronic inflammatory conditions. The method comprises administering a therapeutically effective amount of a compound of structural formula I to an individual in need thereof. The method also encompasses administering to the subject a second therapeutic agent in addition to the compound of structural formula (I). The second therapeutic agent is selected from drugs known to be useful in treating a disease or condition afflicted by an individual in need thereof, for example chemotherapeutic agents and/or radiation known to be useful in treating certain cancers.

In some preferred embodiments, ring B is phenyl, naphthyl, pyridinyl, pyridazinyl, pyrazinyl or pyrimidinyl.

In some preferred embodiments, R is

(여기서 p는 0 내지 4임),

,

,

(여기서 q는 0 내지 2임), 및 -(CH₂)_2-4-C₆H₅를 포함하나, 이제 제한되지는 않는다.

(Where p is 0 to 4),

,

,

(Wherein q is from 0 to 2), and -(CH ₂ ) _2-4 -C ₆ H ₅ , but is not limited thereto.

Specific R groups include, but are not limited to:

,

,

,

,

,

및

(여기서 n은 0 또는 1임)이나 이에 제한되지는 않는다.In some preferred embodiments, R ₁ is

,

,

,

,

,

And

(Where n is 0 or 1), but is not limited thereto.

,

,

,

,

,

,

,

,

,

,

,

및

를 포함하나, 이에 제한되지는 않는다.The specific R ₁ group

,

,

,

,

,

,

,

,

,

,

,

And

Including, but is not limited to.

이고 Y는 -NH-이다.In some preferred embodiments, X is

And Y is -NH-.

In another preferred embodiment, X is SO ₂ and Y is absent.

이고 Y는 부존재이다.In another preferred embodiment, X is

And Y is absent.

이고 Y는 -NH-이다.In another preferred embodiment, X is

And Y is -NH-.

이고 Y는 -O-이다.In another preferred embodiment, X and X'are

And Y is -O-.

Additionally, salts, hydrates, solvates and prodrugs of the compounds of the present invention are also included in the present invention and can be used in the methods disclosed herein. The invention further includes all possible stereoisomers and geometric isomers of the compounds of structural formula (I). The present invention includes both racemic compounds and optically active isomers. When a compound of structural formula (I) is desired as a single enantiomer, it can be obtained by decomposition of the final product, or by stereospecific synthesis from isomerically pure starting materials or by the use of chiral auxiliary reagents, e.g. See, for example, [Z. See Ma et al., Tetrahedron: Asymmetry, 8(6), pages 883-888 (1997). Decomposition of the final product, intermediate or starting material can be accomplished by any suitable method known in the art. Additionally, where tautomers of compounds of structural formula (I) are possible, the present invention is intended to include all tautomeric forms of the compounds.

The compounds of the present invention may exist as salts. Pharmaceutically acceptable salts of the compounds of the present invention are often preferred in the methods of the present invention. As used herein, the term “pharmaceutically acceptable salt” is obtained by reaction with a physiologically acceptable compound of the present invention (eg, with an acid or base) in a target animal (eg, a mammal). To become). Salts of the compounds of the present invention may be derived from inorganic or organic acids and bases. The term “pharmaceutically acceptable salt” also refers to the zwitterionic form of the compound of structural formula (I). Salts of the compounds of formula I may be prepared during the final isolation and purification of the compound, or may be prepared individually by reacting the compounds with an acid having a suitable cation. Pharmaceutically acceptable salts of the compounds of structural formula I may be acid addition salts formed with pharmaceutically acceptable acids. Examples of acids that can be used to form pharmaceutically acceptable salts include inorganic acids such as nitric acid, boric acid, hydrochloric acid, hydrobromic acid, sulfuric acid and phosphoric acid, and organic acids such as oxalic acid, maleic acid, succinic acid and citric acid. . Non-limiting examples of salts of the compounds of the present invention are hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, 2-hydroxyethanesulfonate, phosphate, hydrogen phosphate, acetate, adipate, alginate, as Partate, benzoate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, glycerol phosphate, hemisulphate, heptanoate, hexanoate, formate, succinate, fumarate, maleate, Ascorbate, isethionate, salicylate, methanesulfonate, mesitylenesulfonate, naphthalenesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate, 3-phenyl Propionate, picrate, pivalate, propionate, trichloroacetate, trifluoroacetate, phosphate, glutamate, bicarbonate, paratoluenesulfonate, undecanoate, lactate, citrate, tartrate , Gluconate, methanesulfonate, ethandisulfonate, benzene sulfonate and p-toluenesulfonate salts, but are not limited thereto. Examples of bases include alkali metal (e.g., sodium) hydroxide, alkaline earth metal (e.g., magnesium) hydroxide, ammonia, and compounds of the formula NW ₄ ^{+ wherein} _{W is C 1-4} alkyl, and the like. However, it is not limited thereto. Also available amino groups present in the compounds of the present invention include methyl, ethyl, propyl and butyl chloride, bromide and iodide; Dimethyl, diethyl, dibutyl and diamyl sulfate; Decyl, lauryl, myristyl and steryl chlorides, bromide and iodide; And benzyl and phenethyl bromide.

의 혼합물 둘 다를 포함한다.Compounds of structural formula (I) may contain one or more asymmetric centers and thus may exist as stereoisomers. The present invention includes both mixtures and individual stereoisomers. In particular, the compounds of structural formula I are the individual cis- and trans-isomers, and cis- and trans-isomers, for example

Includes both mixtures of.

As used herein, the term “prodrug” is either released within a target physiological system (eg, enzymatically, physiologically, mechanically, electromagnetically) or in which the prodrug is converted into an active drug (eg, spontaneously. Or enzymatic) biotransformation. Prodrugs are designed to overcome problems associated with stability, toxicity, lack of specificity or limited bioavailability.

Prodrugs often provide the advantage of solubility, tissue compatibility or delayed release in mammalian organisms. (See, eg, Bundgard, “Design of Prodrugs”, pp. 7-9, 21-24, Elsevier, Amsterdam (1985); And Silverman, “The Organic Chemistry of Drug Design and Drug Action”, pp. 352 -401, Academic Press, San Diego, CA (1992)). Exemplary prodrugs include the active drug molecule itself and a chemical masking group (eg, a group that reversibly inhibits drug activity). Some preferred prodrugs are variants or derivatives of compounds having groups that are cleavable under metabolic conditions. Exemplary prodrugs are in vivo or in vitro when they undergo solvolysis under physiological conditions or undergo enzymatic degradation or other biochemical modifications (e.g., phosphorylation, hydrogenation, dehydrogenation, glycosylation). Becomes pharmaceutically active in Typical prodrugs are acid derivatives, such as esters prepared by reaction of a parent acid with a suitable alcohol (e.g., lower alkanols), amides prepared by reaction of a parent acid compound with an amine, or an acylated base derivative ( For example, lower alkylamides).

Specific compounds of the present invention include, but are not limited to, compounds having the structures described below.

The present invention provides inhibitors of IAP proteins as exemplified by compounds of structural formula I for the treatment of various diseases and conditions in which inhibition of IAP proteins has a beneficial effect. In one embodiment, the invention comprises administering a therapeutically effective amount of a compound of structural formula (I) to an individual suffering from the disease or condition for which inhibition of the IAP protein benefits. It is about how to cure.

IAP protein inhibitors of the present invention when administered as monotherapy to induce apoptosis in cancer cells dependent on IAP function, or compared to the corresponding proportion of cells in animals treated with cancer treatment drugs or radiation therapy alone. When administered in a temporal relationship with other anti-cancer therapies to further increase the proportion of cancer cells susceptible to performing an apoptosis program, it meets the need for treatment of multiple cancer types.

As used herein, the term “anti-cancer therapy” refers to therapeutic agents (eg, chemotherapeutic compounds and/or molecular therapeutic compounds), radiation therapy and surgical interventions used in the treatment of hyperproliferative diseases such as cancer in a mammal. .

The method of the present invention can be achieved by administering a compound of structural formula (I) as a pure compound or as a pharmaceutical composition. Administration of the pharmaceutical composition or the pure structural compound of formula I can be carried out during or after the onset of the disease or condition of interest. Typically, pharmaceutical compositions are sterile and do not contain any toxic, carcinogenic or mutagenic compounds that will cause adverse reactions upon administration. A second therapeutic agent useful for the treatment of diseases and conditions for which inhibition of the IAP protein benefits, and optionally, individually or packaged together, a compound of structural formula I, and an insert containing instructions for the use of these actives. Additional kits are provided.

In many embodiments, the compound of structural formula (I) is administered in combination with a second therapeutic agent useful in the treatment of a disease or condition for which inhibition of the IAP protein benefits. The second therapeutic agent is different from the compound of structural formula (I). The compound of structural formula (I) and the second therapeutic agent can be administered simultaneously or sequentially to achieve the desired effect. In addition, the compound of structural formula I and the second therapeutic agent can be administered from a single composition or from two separate compositions.

The second therapeutic agent is administered in an amount that provides its desired therapeutic effect. Ranges of effective dosages for each second therapeutic agent are known in the art, and the second therapeutic agent is administered to an individual in need thereof within this established range.

In certain embodiments, the combination treatment comprising administering a therapeutically effective amount of a compound of structural formula (I) and a second therapeutic agent is a greater tumor response and greater clinical response compared to treatment with a compound of structural formula (I) or a second therapeutic agent alone. Generate enemy benefits.

It is also possible to achieve the administration of a lower dose of a second therapeutic agent using the compound of structural formula I, thus achieving lower toxicity and better tolerability, and the same tumor response as the conventional dose of the second therapeutic agent. /Can generate clinical benefit. In addition, because the compounds of the present invention act at least in part by inhibiting the IAP protein, exposure of cancer and supporting cells to a therapeutically effective amount of an IAP protein inhibitor of the present invention causes the cells to react to a second therapeutic agent to initiate an apoptosis program. It can be temporally linked with the attempt to execute. Thus, in some embodiments, administering a compound of the present invention in relation to a second therapeutic agent in a specific temporal relationship provides a particularly effective therapeutic outcome.

The compound of structural formula I and the second therapeutic agent can thus be administered together as a single-unit dose or separately as a multi-unit dose, wherein the compound of structural formula I is administered before the second therapeutic agent or vice versa. One or more doses of the compound of structural formula I and/or one or more doses of a second therapeutic agent may be administered. The compounds of structural formula (I) can thus be used in combination with one or more second therapeutic agents, such as, but not limited to, anticancer agents.

Diseases and conditions that can be treated according to the present invention include, for example, cancer. Bladder carcinoma (including accelerated and metastatic bladder cancer), breast carcinoma, colon carcinoma (including colorectal cancer), kidney carcinoma, liver carcinoma, lung carcinoma (including bovine and non-small cell lung cancer and lung adenocarcinoma), ovarian carcinoma, prostate carcinoma, testicular carcinoma, Genitourinary tract carcinoma, lymphatic carcinoma, rectal carcinoma, laryngeal carcinoma, pancreatic carcinoma (including exocrine pancreatic carcinoma), esophageal carcinoma, gastric carcinoma, gallbladder carcinoma, cervical carcinoma, thyroid carcinoma, kidney carcinoma, and skin carcinoma (including squamous cell carcinoma) Carcinoma, including; Hematopoietic tumors of the lymphatic system, including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma, histocytic lymphoma and Burkitt's lymphoma , Acute and chronic myelogenous leukemia, myelodysplastic syndrome, myeloid leukemia and hematopoietic tumors of the myeloid system including promyelocytic leukemia; Tumors of the central and peripheral nervous system including astrocytoma, neuroblastoma, glioma and Schwanncytoma; Tumors of mesenchymal origin including fibrosarcoma, rhabdomyosarcoma and osteosarcoma; Other tumors, including melanoma, scleroderma, keratoblastoma, seminoma, thyroid follicular cancer, teratocarcinoma, renal cell carcinoma (RCC), pancreatic cancer, myeloma, myeloid and lymphoblastic leukemia, neuroblastoma and glioblastoma. A variety of cancers, including but not limited to, can be treated.

Additional forms of cancer that can be treated with the IAP protein inhibitors of the invention include, for example, adult and pediatric oncology, solid tumor/malignant tumor growth, mucinous and round cell carcinoma, locally advanced tumor, metastatic cancer, Ewing's sarcoma. Including human soft tissue sarcoma, cancer metastasis including lymphatic metastasis, squamous cell carcinoma, especially head and neck carcinoma, esophageal squamous cell carcinoma, oral carcinoma, blood cell malignancies including multiple myeloma, acute lymphocytic leukemia, acute nonlymphocyte leukemia, Leukemia including chronic lymphocytic leukemia, chronic myeloid leukemia and hairy cell leukemia, effusion lymphoma (celiac lymphoma), thymic lymphoma lung cancer (small cell carcinoma, cutaneous T cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, adrenal cortex Cancer, ACTH-producing tumors, non-small cell cancer, breast cancer (including small cell carcinoma) and ductal carcinoma), gastrointestinal cancer (including gastric cancer, colon cancer, colorectal cancer, and polyps associated with colorectal neoplasms), pancreatic cancer, liver cancer, urinary tract Cancer (including bladder cancer, such as primary superficial bladder tumor, invasive transitional cell carcinoma of the bladder and muscle-invasive bladder cancer), prostate cancer, malignant tumors of the female reproductive tract (ovarian carcinoma, primary peritoneal epithelial neoplasm, cervical carcinoma, endometrial Cancer, vaginal cancer, vulvar cancer, uterine cancer and ovarian follicle solid tumors), malignancy of the male reproductive tract (including testicular cancer and penile cancer), kidney cancer (including renal cell carcinoma), brain cancer (intrinsic brain tumor, neuroblastoma, Astrocyte brain tumor, glioma, and metastatic tumor cell invasion in the central nervous system), bone cancer (including osteoma and osteosarcoma), skin cancer (including malignant melanoma, tumor progression of human cutaneous keratinocytes, and squamous cell carcinoma), thyroid cancer, Retinoblastoma, neuroblastoma, peritoneal effusion, malignant pleural effusion, mesothelioma, Wilms' tumor, gallbladder cancer, trophoblastic neoplasm, hemangiopericytoma, and Kaposi's sarcoma.

Another embodiment of the invention is to induce apoptosis and increase the potency of inducing apoptosis in response to an apoptosis inducing signal by the use of IAP protein inhibition of structural formula I. IAP protein inhibitors of the invention also sensitize cells to such inducers, including cells that are resistant to apoptosis inducers. IAP protein inhibitors of the present invention can be used to induce apoptosis in any disorder that can be treated, ameliorated or prevented by induction of apoptosis. Accordingly, the present invention provides a composition and method for treating an animal characterized by overexpressing an IAP protein. In some embodiments, the cells (eg, cancer cells) display an elevated level of expression of the IAP protein compared to a non-pathological sample (eg, a non-cancerous cell). In another embodiment, the cell operably displays an elevated level of expression of the IAP protein by executing an apoptosis program and killing it in response to a therapeutically effective amount of a compound of structural formula I, wherein the response is at least in part Occurs due to dependence on IAP protein function for survival.

In another embodiment, the invention relates to modulating an apoptosis-associated condition associated with one or more apoptosis-modulating agents. Examples of apoptosis-modulating agents include Fas/CD95, TRAMP, TNF RI, DR1, DR2, DR3, DR4, DR5, DR6, FADD, RIP, TNFα, Fas ligand, TRAIL, TRAIL-R1 or antibody against TRAIL-R2, Bcl -2, p53, BAX, BAD, Akt, CAD, PI3 kinase, PP1 and caspase proteins. Other agents involved in the initiation, determination and degradation steps of apoptosis are also included. Examples of apoptosis-modulating agents include agents whose activity, presence, or concentration in a subject can modulate apoptosis. Preferred apoptosis-modulating agents are inducers of apoptosis, such as TNF or TNF-related ligands, in particular TRAMP ligands, Fas/CD95 ligands, TNFR-1 ligands or TRAIL.

These therapies can be used in a variety of settings for the treatment of a variety of cancers. In specific embodiments, the individual in need of treatment has previously undergone cancer treatment. Such prior therapy includes, but is not limited to, prior chemotherapy, radiotherapy, surgery or immunotherapy, such as cancer vaccines.

In one embodiment, the invention provides a method comprising: (a) administering a therapeutically effective amount of an IAP protein inhibitor of structural formula (I) to an individual in need thereof; And (b) administering to the subject a therapeutically effective amount of one or more of radiotherapy, chemotherapy and immunotherapy. The amount administered is each effective in treating cancer. In another embodiment, the amounts are effective together in treating cancer.

In another embodiment, the invention provides a method of treating cancer comprising administering to a subject in need thereof a pharmaceutical composition comprising an IAP protein inhibitor of structural formula I.

In another embodiment, an IAP protein inhibitor of the invention comprises T and B cell mediated autoimmune diseases; Inflammatory disease; infection; Hyperproliferative disease; AIDS; Denatured state; It is used in a method of treating vascular diseases, etc. In some embodiments, infections suitable for treatment with the compositions and methods of the invention include, but are not limited to, infections caused by viruses, bacteria, fungi, mycoplasma, prions, and the like.

The compounds and methods of the present invention are also useful for the treatment of autoimmune disorders or chronic inflammatory conditions. As used herein, the term “autoimmune disorder” refers to any condition in which an organism produces antibodies or immune cells that recognize molecules, cells or tissues of the organism itself. Non-limiting examples of autoimmune disorders include autoimmune hemolytic anemia, autoimmune hepatitis, Berger's disease or IgA nephropathy, celiac sprue, chronic fatigue syndrome, Crohn's disease, dermatitis, fibromyalgia, graft versus host disease, Graves disease, Hashimoto's thyroiditis, idiopathic thrombocytopenia purpura, lichen planus, multiple sclerosis, myasthenia gravis, psoriasis, rheumatic fever, rheumatoid arthritis, scleroderma, Sjogren's syndrome, systemic lupus erythematosus, type 1 diabetes, ulcerative colitis, vitiligo and others. Includes.

Additional diseases and conditions, including cancer, that can be treated by administration of an IAP protein inhibitor of the present invention are disclosed in US Pat. No. 7,960,372; It is incorporated herein by reference in its entirety.

In this method, a therapeutically effective amount of one or more Compound I, typically formulated according to pharmaceutical practice, is administered to a human in need thereof. Whether such treatment is recommended depends on the individual case and is medically evaluated (diagnosed) taking into account the signs present, symptoms and/or dysfunction, the specific signs, risk of developing symptoms and/or dysfunction, and other factors. .

The compounds of structural formula (I) can be prepared by any suitable route, for example oral, buccal, inhalation, sublingual, rectal, vaginal, intracisternal or intrathecal, transurethral, nasal, transdermal, i.e. transdermal or parenteral. Oral (intravenous, intramuscular, subcutaneous, intracoronary, intradermal, intramammary, intraperitoneal, intraarticular, intrathecal, posterior, intrapulmonary injection and/or surgical implantation at a specific site) can be administered. . Parenteral administration can be performed using needles and syringes or using high pressure techniques.

Pharmaceutical compositions include pharmaceutical compositions in which the compound of structural formula (I) is administered in an amount effective to achieve its intended purpose. The exact formulation, route of administration and dosage are determined by taking into account the condition or disease diagnosed by the individual physician. The dosage and interval can be adjusted individually to provide a level of the compound of structural formula I sufficient to maintain the therapeutic effect.

The toxicity and therapeutic efficacy of a compound of structural formula (I) can be determined by standard pharmaceutical procedures in cell culture or laboratory animals, e.g. to determine the maximum tolerated dose (MTD) of the compound, which is defined as the maximum dose that does not cause any toxicity in the animal. Can be determined. The dose ratio between the maximum tolerated dose and the therapeutic effect (eg inhibition of tumor growth) is the therapeutic index. The dosage may vary within this range depending on the dosage form used and the route of administration used. Determination of a therapeutically effective amount is well within the capabilities of one of ordinary skill in the art, particularly in light of the detailed disclosure provided herein.

Structure required for use in therapy The therapeutically effective amount of a compound of formula I depends on the nature of the condition to be treated, the length of time for which activity is desired, and the age and condition of the patient, and is ultimately determined by the practitioner. Dosage and interval can be adjusted individually to provide plasma levels of the IAP protein inhibitor sufficient to maintain the desired therapeutic effect. The desired dose may be conveniently administered as a single dose or as multiple doses administered at appropriate intervals, for example in divided doses of 1, 2, 3, 4 or more times per day. Multiple doses are often desired or required. For example, the IAP protein inhibitors of the present invention can be administered in 4 doses (q4d x 4) delivered as a single dose per day at 4-day intervals; 4 doses delivered as 1 dose per day at 3 day intervals (q3d x 4); One dose delivered per day at 5-day intervals (qd x 5); 1 dose per week for 3 weeks (qwk3); 5 daily doses, 2 days off, and an additional 5 daily doses (5/2/5); Or at the frequency of any dosage regimen determined to be appropriate for the situation.

The compounds of structural formula I used in the methods of the present invention may be administered in an amount of about 0.005 to about 500 milligrams per dose, about 0.05 to about 250 milligrams per dose, or about 0.5 to about 100 milligrams per dose. For example, compounds of structural formula (I) may contain about 0.005, 0.05, 0.5, 5, 10, 20, 30, 40, 50, 100, 150, 200, 250, or about 0.005, 0.05, 0.5, 5, 10, 20, 30, 40, 50, 100, 150, 200, 250 per dose, including all doses between 0.005 and 500 milligrams. It can be administered in an amount of 300, 350, 400, 450 or 500 milligrams.

The dosage of the composition containing the IAP protein inhibitor of structural formula (I), or the composition containing the same, is from about 1 ng/kg to about 200 mg/kg, from about 1 μg/kg to about 100 mg/kg, or about 1 mg/kg. kg to about 50 mg/kg. The dosage of the composition may be any dosage including, but not limited to, about 1 μg/kg. The dosage of the composition is about 1 μg/kg, 10 μg/kg, 25 μg/kg, 50 μg/kg, 75 μg/kg, 100 μg/kg, 125 μg/kg, 150 μg/kg, 175 μg/kg , 200 μg/kg, 225 μg/kg, 250 μg/kg, 275 μg/kg, 300 μg/kg, 325 μg/kg, 350 μg/kg, 375 μg/kg, 400 μg/kg, 425 μg/kg , 450 μg/kg, 475 μg/kg, 500 μg/kg, 525 μg/kg, 550 μg/kg, 575 μg/kg, 600 μg/kg, 625 μg/kg, 650 μg/kg, 675 μg/kg , 700 μg/kg, 725 μg/kg, 750 μg/kg, 775 μg/kg, 800 μg/kg, 825 μg/kg, 850 μg/kg, 875 μg/kg, 900 μg/kg, 925 μg/kg , 950 μg/kg, 975 μg/kg, 1 mg/kg, 5 mg/kg, 10 mg/kg, 15 mg/kg, 20 mg/kg, 25 mg/kg, 30 mg/kg, 35 mg/kg , 40 mg/kg, 45 mg/kg, 50 mg/kg, 60 mg/kg, 70 mg/kg, 80 mg/kg, 90 mg/kg, 100 mg/kg, 125 mg/kg, 150 mg/kg , 175 mg/kg or 200 mg/kg. The above dosage is an example of an average case, but there may be individual cases in which higher or lower dosages are advantageous, and this is within the scope of the present invention. In fact, the physician determines the most appropriate actual dosage regimen for an individual patient, which may vary depending on the age, weight and response of a particular patient.

In the treatment of cancer, the compounds of structural formula I can be administered in combination with chemotherapeutic and/or immunotherapy and/or radiation or in combination with another treatment technique, such as surgery. As used herein, the term chemotherapeutic agents includes anticancer agents, anti-neoplastic agents, and apoptosis-modulating agents.

Embodiments of the present invention include gamma-radiation (10 ^-20 to 10 ^-13 m), X-radiation (10 ^-12 to 10 ^-9 m), ultraviolet (10 nm to 400 nm), visible light (400 nm to 700 nm), infrared radiation (700 nm to 1 mm), and microwave radiation (1 mm to 30 cm) of electromagnetic radiation are used.

Currently, many cancer treatment protocols use radiosensitizers that are activated by electromagnetic radiation, for example X-rays. Examples of X-ray-activated radiosensitizers include metronidazole, misonidazole, desmethylmisonidazole, pimonidazole, ethanidazole, nimorazole, mitomycin C, RSU 1069, SR 4233, EO9, RB 6145, nicotinamide. , 5-bromodeoxyuridine (BUdR), 5-iododeoxyuridine (IUdR), bromodeoxycytidine, fluorodeoxyuridine (FUdR), hydroxyurea, cis-platin and treatment thereof Phase effective analogs and derivatives are included, but are not limited thereto.

Photodynamic therapy (PDT) for cancer uses visible light as a radiation activator for sensitizers. Examples of photodynamic radiosensitizers include hematoporphyrin derivatives, PHOTOFRIN®, benzoporphyrin derivatives, NPe6, tin ethioporphyrin (SnET2), pheophorvid-a, bacteriochlorophyll-a, naphthalocyanine, phthalocyanine, Zinc phthalocyanine and therapeutically effective analogs and derivatives thereof.

Radiosensitizers may be administered with a therapeutically effective amount of one or more compounds in addition to the IAP protein inhibitors of the present invention, such compounds being compounds that promote incorporation of radiosensitizers into target cells, therapeutic agents into target cells, nutrients, and /Or compounds that control the flow of oxygen, chemotherapeutic agents that act on the tumor in the presence or absence of additional radiation, or other therapeutically effective compounds for treating cancer or other diseases. Examples of additional therapeutic agents that can be used with radiosensitizers include 5-fluorouracil (5-FU), leucovorin, oxygen, carbogen, red blood cell transfusion, perfluorocarbons (e.g., FLUOSOLW )®-DA), 2,3-DPG, BW12C, calcium channel blockers, pentoxifylline, anti-angiogenic compounds, hydralazine and L-BSO.

The chemotherapeutic agent can be any pharmacological agent or compound that induces apoptosis. The pharmacological agent or compound can be, for example, a small organic molecule, a peptide, a polypeptide, a nucleic acid or an antibody. Chemotherapeutic agents that can be used include, but are not limited to, alkylating agents, antimetabolites, hormones and antagonists thereof, natural products and derivatives thereof, radioisotopes, antibodies, as well as natural products and combinations thereof. For example, the IAP protein inhibitors of the invention are antibiotics such as doxorubicin and other anthracycline analogs, nitrogen mustards such as cyclophosphamide, pyrimidine analogs such as 5-fluorouracil, cis-platin, hydroxyurea. , Taxol and its natural and synthetic derivatives, and the like. As another example, in the case of a mixed tumor, such as adenocarcinoma of the breast, the tumor comprises gonadotropin-dependent and gonadotropin-independent cells, the compound is leuprolide or goserelin (synthesis of LH-RH Peptide analogs). Other anti-neoplastic protocols include the use of inhibitor compounds in conjunction with another treatment modality, also referred to herein as an "adjuvant anti-neoplastic modality", such as surgery or radiation. Additional chemotherapeutic agents useful in the present invention include hormones and antagonists thereof, radioisotopes, antibodies, natural products and combinations thereof.

Examples of chemotherapeutic agents useful in the methods of the present invention are listed in the table below.

<Table 1>

Microtubule invading agents interfere with mitosis of cells, and their cytotoxic activity is well known in the art. Microtubule invasion agents useful in the present invention are allocolchycin (NSC 406042), halichondrin B (NSC 609395), colchicine (NSC 757), colchicine derivatives (e.g., NSC 33410), dolastatin 10 (NSC 376128), Maytansine (NSC 153858), Rizoxine (NSC 332598), paclitaxel (NSC 125973), TAXOL® derivatives (e.g. NSC 608832), thiocholxine (NSC 361792), trityl cysteine (NSC 83265), Natural and synthetic epothilones including, but not limited to, vinblastine sulfate (NSC 49842), vincristine sulfate (NSC 67574), epothilone A, epothilone B, and discodermolide (Service, (1996) Science, 274:2009]), including, but not limited to, estramustine, nocodazole, MAP4, and the like. Examples of such agents are also described in Bulinski (1997) J. Cell Sci. 110:3055 3064; Panda (1997) Proc. Natl. Acad. Sci. USA 94:10560-10564; Muhlradt (1997) Cancer Res. 57:3344-3346; Nicolaou (1997) Nature 397:268-272; Vasquez (1997) Mol. Biol. Cell. 8:973-985; And Panda (1996) J. Biol. Chem. 271:29807-29812.

Cell proliferation inhibitors that can be used are hormones and steroids (including synthetic analogues): 17-α-ethynylestradiol, diethylstilvestrol, testosterone, prednisone, fluoxymesterone, dromostanolone propionate, testostelactone. , Megestrolacetate, methylprednisolone, methyl-testosterone, prednisolone, triamcinolone, chlorotrianicene, hydroxyprogesterone, aminoglutethymide, estramustine, medroxyprogesterone acetate, leuprolide, flutamide, toremifene, Zoladex is included, but is not limited thereto.

Other cytostatic agents are anti-angiogenic agents such as matrix metalloproteinase inhibitors and other VEGF inhibitors such as anti-VEGF antibodies and small molecules such as ZD6474 and SU668. Anti-Her2 antibodies can also be used. The EGFR inhibitor is EKB-569 (irreversible inhibitor). Also included is antibody C225 immunospecific for EGFR and Src inhibitors.

Also suitable for use as a cytostatic agent is CASODEX® (Bicalutamide, Astra Zeneca), which renders androgen-dependent carcinoma non-proliferative. Another example of a cytostatic agent is the antiestrogen tamoxifen (TAMOXIFEN)®, which inhibits the proliferation or growth of estrogen dependent breast cancer. Inhibitors of transmission of cell proliferation signals are cytostatic agents. Representative examples include epidermal growth factor inhibitors, Her-2 inhibitors, MEK-1 kinase inhibitors, MAPK kinase inhibitors, PI3 inhibitors, Src kinase inhibitors and PDGF inhibitors.

Antimicrobial therapeutic agents can also be used as the second therapeutic agent in the present invention. Any agent that is capable of killing, inhibiting or otherwise attenuating the function of the microbial organism, as well as any agent contemplated to have such activity may be used. Antimicrobial agents include, but are not limited to, natural and synthetic antibiotics, antibodies, inhibitory proteins (eg, defensins), antisense nucleic acids, membrane disruptors, and the like, and are used alone or in combination. Indeed, any type of antibiotic may be used including, but not limited to, antibacterial agents, antiviral agents, antifungal agents, and the like.

Additional second therapeutic agents that can be administered in combination with the IAP protein inhibitors of the present invention are disclosed in US Pat. No. 7,960,372, which is incorporated herein by reference in its entirety.

The compounds of the present invention are typically administered in admixture with a pharmaceutical carrier selected with respect to the intended route of administration and standard pharmaceutical practice. Pharmaceutical compositions for use in accordance with the present invention are formulated in a conventional manner using one or more physiologically acceptable carriers comprising excipients and adjuvants which facilitate processing of the compounds of structure I.

These pharmaceutical compositions can be prepared, for example, by conventional mixing, dissolving, granulating, dragee-making, emulsifying, encapsulating, entrapping or lyophilizing processes. Proper formulation depends on the route of administration chosen. When a therapeutically effective amount of a compound of structural formula (I) is administered orally, the composition is typically in the form of tablets, capsules, powders, solutions or elixirs. When administered in the form of a tablet, the composition may further contain a solid carrier such as gelatin or adjuvant. Tablets, capsules and powders contain from about 0.01% to about 95%, and preferably from about 1% to about 50%, of a compound of structural formula (I). When administered in liquid form, liquid carriers such as water, petroleum, or oils of animal or plant origin may be added. The liquid form of the composition may further contain a physiological saline solution, dextrose or other saccharide solution or glycol. When administered in liquid form, the composition contains from about 0.1% to about 90% by weight, and preferably from about 1% to about 50% by weight of a compound of structural formula (I).

When a therapeutically effective amount of a compound of structural formula (I) is administered by intravenous, dermal or subcutaneous injection, the composition is in the form of a pyrogen-free, parenterally acceptable aqueous solution. Preparation of such a parenterally acceptable solution, taking full consideration of pH, isotonicity, stability, etc., is within the scope of the related art. Compositions preferred for intravenous, dermal or subcutaneous injection typically contain an isotonic vehicle.

The compounds of structural formula (I) can be readily combined with pharmaceutically acceptable carriers well known in the art. Such a carrier allows the active agent to be formulated into tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc. for oral intake by the patient to be treated. Pharmaceutical preparations for oral use are by adding a compound of structural formula (I) to a solid excipient, optionally grinding the resulting mixture, and processing the granule mixture after adding suitable auxiliaries if desired to obtain tablets or dragee cores. Can be obtained. Suitable excipients include, for example, fillers and cellulose preparations. If desired, a disintegrant can be added.

The compounds of structural formula (I) can be formulated for parenteral administration by injection, for example by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, for example in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulation agents such as suspending, stabilizing and/or dispersing agents.

Pharmaceutical compositions for parenteral administration contain an aqueous solution of the active agent in water-soluble form. In addition, suspensions of compounds of structural formula I can be prepared as suitable oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils or synthetic fatty acid esters. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the compound and allow the preparation of highly concentrated solutions. Alternatively, the compositions of the present invention may be present in powder form consisting of a suitable vehicle, for example sterile pyrogen-free water, prior to use.

The compounds of structural formula (I) can also be formulated as rectal compositions, such as suppositories or retention enemas, for example containing conventional suppository bases. In addition to the formulations previously described, the compounds of structural formula I can also be formulated as depot formulations. Such long acting agents can be administered by implantation (eg, subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, compounds of the structural formula (I) can be formulated with suitable polymeric or hydrophobic materials (eg, as emulsions in acceptable oils) or ion exchange resins.

In particular, the compounds of the structural formula (I) are orally in the form of tablets containing excipients, such as starch or lactose, or in the form of capsules or bulos alone or mixed with excipients, or elixirs or suspensions containing flavoring or coloring agents. , Can be administered buccally or sublingually. Such liquid formulations can be prepared with pharmaceutically acceptable additives such as suspending agents. The compounds of structural formula I can also be injected parenterally, for example intravenously, intramuscularly, subcutaneously or intracoronally. For parenteral administration, IAP protein inhibitors are optimally used in the form of sterile aqueous solutions, which may contain salts or monosaccharides such as mannitol or glucose, such as salts or other substances that render the solution isotonic with blood.

As a further embodiment, the invention includes kits comprising one or more compounds or compositions packaged in a manner that facilitates their use in carrying out the methods of the invention. In one simple embodiment, the kit is packaged in a container, such as a sealed bottle or tube, as useful in the practice of the method, comprising a compound or composition described herein (e.g., a compound of structure I and an optional second therapeutic agent. Composition), and a label describing the use of the compound or composition for carrying out the method of the present invention is affixed to the container or included in the kit. Preferably, the compound or composition is packaged in unit dosage form. The kit may further comprise a device suitable for administration of the composition according to the intended route of administration.

Previous IAP protein inhibitors had properties that hindered their development as therapeutics. According to an important feature of the present invention, compounds of structural formula I have been synthesized and evaluated as inhibitors of IAP proteins. _{For example, the compounds of the present invention typically have a binding affinity (IC 50} ) for an IAP protein of less than 100 nM, less than 50 nM, less than 25 nM and less than 10 nM.

Compound synthesis

The compound of the present invention was prepared as follows. The following synthetic scheme represents the reaction used to synthesize compounds of structural formula (I). Modifications and alternative schemes for preparing the IAP protein inhibitors of the present invention are within the capabilities of those skilled in the art.

Solvents and reagents were obtained commercially and used without further purification. The chemical shift (δ) of the NMR spectrum was reported as the δ value (ppm) of the downfield relative to the internal standard, with the multiplicity reported in the conventional manner.

All temperatures are in degrees Celsius unless otherwise stated.

In synthetic methods and examples, and throughout the specification, abbreviations have the following meanings.

<Synthesis Scheme 1>

Except for having a cyclopropyl ring in R, each compound of structural formula (I) is synthesized according to the method set forth in Synthesis Scheme 1 above. Compound 2 is described in [Q. Cai et al., J. Med. Chem., 2011, 2714-26]. Protection of the amino group by Cbz in compound 2 provided carbamate 3. Hydrolysis of the methyl ester in carbamate 3 gave acid 4. Condensation of acid 4 with a series of amines gave each amide 5. Removal of the Boc protecting group in amide 5 gave amine 6. Condensation of amine 6 and L-N-Boc-N-methyl-alanine gave amide 7. Cleavage of the Cbz protecting group in amide 7 gave amine 8.

Condensation of amine 8 with a series of diisocyanates (9), and subsequent removal of the Boc protecting group, resulted in a bis-urea containing the Smac mimetic. Condensation of amine 8 with a series of diisothiocyanates (10) and subsequent removal of the Boc protecting group yielded a bis-thiourea containing the Smac mimetic. Condensation of amine 8 with a series of dicarbonochloridate (12) and subsequent removal of the Boc protecting group yielded a bis-carbamate containing the Smac mimetic. Condensation of amine 8 with a series of disulfonyl chlorides and subsequent removal of the Boc protecting group resulted in bis-sulfonamides containing Smac mimetics.

<Synthesis Scheme 2>

The synthesis of a compound of structural formula I having a cyclopropyl ring in R is shown in Synthesis Scheme 2 above. Condensation of compound 2 with diisocyanate, diisothiocyanate, dicarbonylchloridate or disulfonyl chloride each provided intermediate 13. Removal of the Boc protecting group in compound 13, and subsequent condensation with L-N-Boc-N-methyl-Ala gave amide 14. Hydrolysis of the methyl ester in amide 14 gave a series of acids. Condensation of an acid with a series of amines and subsequent deprotection of the Boc protecting group gave the final compound.

Binding affinity for XIAP linker-BIR2-BIR3, cIAP1-BIR3, and cIAP-2 BIR2

The binding affinity of the compounds of the present invention to the proteins XIAP linker-BIR2-BIR3 (residues 120-356), cIAP1-BIR3 (residues 253-363), and cIAP-2 BIR3 (residues 238-349) is fluorescently polarized (FP ) Based competitive test. For the cIAP-1 BIR3 and cIAP-2 BIR3 assays, a fluorescently labeled Smac mimetic (Smac-2F) was used as a fluorescent probe. _{The K d} values of Smac-2F for cIAP-1 BIR3 and cIAP-2 BIR3 were determined by monitoring the total fluorescence polarization of a mixture consisting of a fixed concentration of a fluorescent probe and an increasing concentration of protein to maximum full saturation. Fluorescence polarization values were determined in a Microfluor 2 96-well, black, round-bottom plate (Thermo Scientific) in an Infinite M-1000 plate reader (Tecan US); Research Triangle Park, NC). In each well, 1 nM of SMAC-2F and increasing concentrations of protein were added in assay buffer (100 mM potassium phosphate containing 4% DMSO, pH 7.5, 100 μg/ml bovine γ-globulin, 0.02% sodium azide, in vitro. Rosen (Invitrogen)) in a final volume of 125 μl. Plates were incubated for 1-2 hours at room temperature and mixed with gentle shaking to ensure equilibrium. Polarization values in millipolarization units (mP) were measured at an excitation wavelength of 485 nm and an emission wavelength of 530 nm. _{Then calculate the equilibrium dissociation constant (K d} ) by fitting the S-type dose-dependent FP increase as a function of protein concentration using Graphpad Prism 5.0 software (Graphpad Software, San Diego, CA). I did.

The K _i value of the compound was determined through a dose-dependent competitive binding experiment in which a serial dilution of the compound competes with a fluorescent probe of a fixed concentration for binding to a protein of a fixed concentration (typically 2 to 2 _{of the determined K d value.} 3 times). 5 μl of test compound in DMSO and 120 μl of preincubated protein/tracer complex in assay buffer (100 mM potassium phosphate, pH 7.5, 100 μg/ml bovine γ-globulin, 0.02% sodium azide, Invitrogen). The mixture was added to the assay plate and incubated at room temperature for 2 hours with gentle shaking. The final concentrations of protein and probe were 3nM and 1nM, 5nM and 1nM for the assays for cIAP-1 BIR3 and cIAP-2 BIR3, respectively. A negative control containing only the protein/probe complex (corresponding to 0% inhibition), and a positive control containing only free probe (corresponding to 100% inhibition) were included in each assay plate. FP values were measured as described above. IC ₅₀ values were determined by nonlinear regression fit of the competition curve. _{The K i} values of the competitive inhibitors were calculated based on the measured IC _{50 values, the K d} values of the probes for different proteins, and the concentration of proteins and probes in the competitive assay, using the previously described derivation equation.

The FP-based assay for the XIAP linker-BIR2-BIR3 protein was performed in the same procedure. In this assay, a divalent fluorescently tagged peptidic Smac mimetic (Smac-1F) was used as a fluorescent probe, and its K _d value for XIAP linker-BIR2-BIR3 was similarly determined through saturation experiments. 0.01% of Triton X-100 was added in the assay buffer to achieve stable fluorescence and polarization values of the dimer fluorescent probe. The final protein and probe concentrations used in the competitive assay were 3nM and 1nM, respectively.

Inhibition of cell growth in MDA-MB-231 breast cancer and SK-OV-3 ovarian cancer cell lines

1 shows the antitumor activity of Example 2 and Example 24 in an MDA-MB-231 xenograft model in nude mice. Treatment was started when the tumor reached an average volume of 80 mm ^3. Example 24 was given intravenously at a weekly dose (qwkx4, iv) of 10 mg/kg for 4 weeks. Example 2 was given at 3 mg/kg once a week for 4 weeks (qwkx4, iv). Control treatment served as vehicle control. Each group consists of 8-10 mice, and each mouse has one tumor. Tumor regression was achieved in the case of Examples 2 and 24.

Claims (11)

(a) container; (b1) a packaged composition comprising a compound having the structural formula (I), or a pharmaceutically acceptable salt, hydrate, or solvate thereof; And (c) cancer, T and B cell mediated autoimmune diseases, inflammatory diseases, infections, hyperproliferative diseases, AIDS, degenerative conditions, vascular diseases, autoimmune hemolytic, comprising a package insert containing instructions for the use of the composition. Anemia, autoimmune hepatitis, Berger's disease or IgA nephropathy, celiac sprue, chronic fatigue syndrome, Crohn's disease, dermatitis, fibromyalgia, graft versus host disease, Graves' disease, Hashimoto's thyroiditis, idiopathic thrombocytopenia purpura, lichen planus, Used in the treatment of a disease or condition selected from the group consisting of multiple sclerosis, myasthenia gravis, psoriasis, rheumatic fever, rheumatoid arthritis, scleroderma, Sjogren's syndrome, systemic lupus erythematosus, type 1 diabetes, ulcerative colitis, and vitiligo. Kit for doing.
<Formula I>

In the above formula, X is

,

,

And -SO ₂ -;
X is

,

, or

When Y is selected from the group consisting of -NH-, -O- and -S-, and when X is -SO ₂ -, Y is absent;
R is

,

(Where ring A is a C _4-8 aliphatic ring),

And

Is selected from the group consisting of, wherein ring B is an aryl or nitrogen atom-containing heteroaryl and ring B is optionally substituted;
R ₁ is -(CH ₂ ) _4-10 -,

,

,

,

,

(Where Z is O, S or NH) and

(Where n is 0, 1 or 2) is selected from the group consisting of, wherein the B ring is an aryl or nitrogen atom-containing heteroaryl

The ring is optionally substituted. The method of claim 1, further comprising (b2) a packaged composition comprising a second therapeutic agent, wherein the second therapeutic agent is a chemotherapeutic agent, and the package insert provides instructions for the use of compositions administered simultaneously or sequentially. The kit that contains. The kit according to claim 1 or 2, wherein the container is a sealed bottle or tube. The kit of claim 1 or 2, wherein the package insert is affixed to the container or contained within the kit. The kit according to claim 1 or 2, further comprising a device suitable for administration of the composition according to the intended route of administration. The kit of claim 5, wherein the intended route of administration is intravenous or subcutaneous injection. The kit of claim 5, wherein the intended route of administration is intravenous injection. The kit of claim 5, wherein the intended route of administration is subcutaneous injection. The kit according to claim 1 or 2, for use in the treatment of cancer. The method of claim 1 or 2, further comprising: T and B cell mediated autoimmune diseases; Inflammatory disease; infection; Hyperproliferative disease; AIDS; Denatured state; And a kit for use in the treatment of a disease or condition selected from the group consisting of vascular diseases. The method of claim 1 or 2, wherein autoimmune hemolytic anemia, autoimmune hepatitis, Berger's disease or IgA nephropathy, celiac sprue, chronic fatigue syndrome, Crohn's disease, dermatitis, fibromyalgia, graft versus host disease, Graves Disease, Hashimoto's thyroiditis, idiopathic thrombocytopenia purpura, lichen planus, multiple sclerosis, myasthenia gravis, psoriasis, rheumatic fever, rheumatoid arthritis, scleroderma, Sjogren's syndrome, systemic lupus erythematosus, type 1 diabetes, ulcerative colitis, and vitiligo Kit for use in the treatment of a disease or condition selected from the group consisting of.

Images